Apoptosis vurses necrosis - Way to find wether the cell death is due to apoptosis or necrosis? (Oct/21/2008 )

I treated cells with certain chemical, as expected cells die. But, I'm curious whether the cell die following apoptitic path or not... Or they die due to toxicity? I"m not sure about that.. is there any way to distinguish between cell apoptosis and necrosis??? Please help me out!!!

-star*-

Caspase 3 or cleaved PARP will do it. If you get positive signal with either of these, it is apoptosis, otherwise it is some other cell death pathway such as necrosis or autophagy.

-bob1-

Hi!

Apart from caspase detection, etc... an easy way to see wether apoptosis or necrosis is ocurring is by flow cytometry detecting Annexin V convolution (apoptosis) or IP or DAPI detection (necrosis).

-Estersan-

QUOTE (star* @ Oct 21 2008, 02:59 AM)

I treated cells with certain chemical, as expected cells die. But, I'm curious whether the cell die following apoptitic path or not... Or they die due to toxicity? I"m not sure about that.. is there any way to distinguish between cell apoptosis and necrosis??? Please help me out!!!

it also exists caspase-3-independent apsptose; to say apoptosos vs necrosis is in some cases difficult to distinguish as a necrosis step can happen during apoptosis...

hard features for apoptosis are membrane blebbing (zeiosis) and disintegration of membrane phospholipids (analysis with annexin V was already suggested), hypercondensation of chromatin, DNA fragmentation, and shrinkage of cells;

a hard fact for necrosis is swelling of cells

-The Bearer-

Flowcytometry with Annexin V and 7AAD or PI should be enough.

-Bungalow Boy-

QUOTE (star* @ Oct 21 2008, 06:59 PM)

I treated cells with certain chemical, as expected cells die. But, I'm curious whether the cell die following apoptitic path or not... Or they die due to toxicity? I"m not sure about that.. is there any way to distinguish between cell apoptosis and necrosis??? Please help me out!!!

How do U determine the cells are dead?

-Bungalow Boy-

QUOTE (Estersan @ Oct 22 2008, 01:32 AM)

Hi!

Apart from caspase detection, etc... an easy way to see wether apoptosis or necrosis is ocurring is by flow cytometry detecting Annexin V convolution (apoptosis) or IP or DAPI detection (necrosis).

IP?....you mean PI?

and how do you expect to differentiate necrosis from apoptosis by DAPI....it is impossible...DAPI stains all of'em....we are staining our dead/live cells by DAPI....so I don't think DAPI staining would solve the problem.

but AO/PI is a different story, never tried it to be honest.

-Curtis-

QUOTE (Curtis @ Oct 26 2008, 02:08 AM)

QUOTE (Estersan @ Oct 22 2008, 01:32 AM)

Hi!

Apart from caspase detection, etc... an easy way to see wether apoptosis or necrosis is ocurring is by flow cytometry detecting Annexin V convolution (apoptosis) or IP or DAPI detection (necrosis).

IP?....you mean PI?

and how do you expect to differentiate necrosis from apoptosis by DAPI....it is impossible...DAPI stains all of'em....we are staining our dead/live cells by DAPI....so I don't think DAPI staining would solve the problem.